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    UNIVERSITI PUTRA MALAYSIA

    TOXICITY EVALAUTIONS OF ETHANOLIC EXTRACT OF Christia

    vespertilionis (L. F.) BAKH. F. IN MALE SPRAGUE DAWLEY RATS

    NURUL SYAHIRAH BINTI AHMAD SAYUTI

    FPV 2018 15

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    PM TOXICITY EVALAUTIONS OF ETHANOLIC EXTRACT OF Christia vespertilionis (L. F.) BAKH. F. IN MALE SPRAGUE DAWLEY RATS

    By

    NURUL SYAHIRAH BINTI AHMAD SAYUTI

    Thesis submitted to the School of Graduate Studies, Universiti Putra Malaysia, in �ulfilment of the �equirement for the Degree of Master �f Science

    January 2018

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    COPYRIGHT

    All material contained within the thesis, including without limitation text, logos, icons,

    photographs and all other artwork, is copyright of Universiti Putra Malaysia unless

    other stated. Use may be made of any material contained within the thesis for non-

    commercial proposes from the copyright holder. Commercial use of material may only

    be made with the express, prior written permission of Universiti Putra Malaysia.

    Copyright © Universiti Putra Malaysia

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    Abstract of thesis presented to Senate of Universiti Putra Malaysia in fulfilment of

    the requirement for the degree of Master of Science

    TOXICITY EVALUATIONS OF ETHANOLIC EXTRACT OF Christia vespertilionis (L.F.) BAKH. F. IN MALE SPRAGUE DAWLEY RATS

    By

    NURUL SYAHIRAH BINTI AHMAD SAYUTI

    January 2018

    Chairman : Associate Professor Hazilawati binti Hamzah, PhD Faculty : Veterinary Medicine

    The term Butterfly tea refers to the decoction of Christia vespertilionis (L.f.) Bakh. f. leaves which is widely consumed by cancer patients throughout Malaysia, and it has

    gained a huge popularity among researchers yearning to discover the real potential of

    this plant. Herein, this study is aimed at evaluating possible toxicity in a 14-day acute,

    28-day subacute and 90-day subchronic oral toxicity of the ethanolic extract Christia vespertilionis (L.f.) Bakh. f. in male Sprague Dawley rats. The 14-day acute toxicity study was conducted to detect lethal dose 50 (LD50) Christia vespertilionis (L.f.) Bakh. f. while the 28-day subacute and 90-day subchronic toxicity studies are to detect the

    non-observed-adverse-effect level (NOAEL). In the acute toxicity study, rats were

    divided into control, 5% DMSO (vehicle) and 2000 mg/kg groups. The extract was

    administered orally on day 1 and observed for 14 days. Meanwhile, in the subacute

    and subchronic toxicity studies, a total of 30 rats were divided into control, 5% DMSO

    (vehicle), low dose (75 mg/kg), medium dose (125 mg/kg) and high dose (250 mg/kg)

    groups. The extract was administered daily from day 1 until day 28 for subacute and

    day 90 for subchronic. Standard toxicology parameters including mortality,

    behavioural changes, motor-neuronal abnormalities, feed-water consumption pattern

    and body weight were measured. The haematological, serum biochemical parameters

    and histopathological assessment of kidney and liver functions were also carried out.

    Results of acute oral toxicity showed that single dose (2000 mg/kg) of ethanol extract

    of Christia vespertilionis (L.f.) Bakh. f. leaves induced no treatment-related signs of toxicity or mortality in male Sprague Dawley rats. The haematological results also

    showed no changes in the control and treated groups in all 3 studies. However, serum

    biochemistry results for acute study, showed a significant decrease in the CK and AST

    level when compared with the control and treated groups. Meanwhile results for serum

    biochemistry in subacute and subchronic showed no changes in the control and treated

    groups for both studies. Organs to body weights ratio after euthanisation in all 3

    studies showed no significant differences when comparing treated and control groups.

    On histopathological analysis, acute study showed significant differences (p

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    lesions observed on hepatic necrosis (mild to moderate) and degeneration (very mild)

    in the treated group (2000 mg/kg). Meanwhile, subacute study showed significant

    differences (p

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    Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia ��������

    meme uhi keperluan� � � ijazah Master Sains

    ANALISIS TOKSISITI DARIPADA EKSTRAK ETANOL DAUN Christia vespertilionis (L.F.) BAKH. F. PADA TIKUS JANTAN SPRAGUE DAWLEY

    Oleh

    NURUL SYAHIRAH BINTI AHMAD SAYUTI

    Januari 2018

    Pengerusi : Profesor Madya Hazilawati binti Hamzah, PhD Fakulti : Perubatan Veterinar

    Teh Rerama merujuk kepada rebusan daun Christia vespertilionis (L.f.) Bakh. f. yang di konsumi oleh pesakit kanser di Malaysia dan telah menjadi popular di Malaysia

    bukan sahaja dikalangan pesakit kanser tetapi juga kepada penyelidik untuk mengkaji

    pontensi sebenar pokok ini. Bagi kajian ini, tujuan kajian ini adalah untuk menilai

    aktiviti toksisiti 14-hari akut, 28-hari subakut dan 90-hari subkronik ekstrak etanol

    Christia vespertilionis (L.f.) Bakh. f. ke atas tikus jantan spesis Sprague Dawley. Akut 14-hari aktiviti toksisiti dijalankan untuk mengenalpasti dos Christia vespertilionis (L.f.) Bakh. f. yang menyebabkan 50% kematian (LD50) dan diikuti kajian 28-hari

    subakut dan subkronik toksisiti untuk mengenalpasti dos yang tidak menyebabkan

    kesan sampingan (NOAEL). Untuk kajian toksisiti akut, tikus dibahagikan kepada

    kumpulan kawalan normal, 5% DMSO (transportasi) dan kumpulan dos 2000mg/kg.

    Ekstrak diberi secara oral pada hari pertama (1) dan diperhati selama 14 hari.

    Manakala, untuk subakut dan subkronik kajian toksisiti, sebanyak 30 ekor tikus

    dibahagikan kepada kumpulan kawalan normal, 5% DMSO (transporatasi), dos

    rendah (75 mg/kg), dos sederhana (125 mg/kg) dan kumpulan dos tinggi (250 mg/kg).

    Ekstrak diberikan setiap hari selama 28 hari subakut dan 90 hari subkronik. Aktiviti

    toksisiti dinilai melalui parameter-parameter seperti kadar kematian, perubahan

    perangai, ketidaknormalan motor dan neuron, berat badan dan polar makanan dan air

    yang dikonsumi. Selain itu, analisis darah, serum biokimia dan histopatologi buah

    pinggang dan hati telah dijalankan. Keputusan untuk kajian akut oral toksisiti

    menunjukkan satu dos 2000mg/kg etanol ekstrak daun Christia vespertilionis (L.f.) Bakh. f. tidak menyebabkan kematian terhadap tikus jantan spesis Sprague Dawley. Keputusan analisis darah untuk ketiga-tiga kajian juga tidak menunjukkan perubahan

    yang ketara di antara kumpulan kawalan normal dan kumpulan yang diberi ekstrak.

    Keputusan analisis serum biokimia dalam kajian akut toksisiti menunjukkan paras CK

    dan AST signifikan (p

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    kumpulan kawalan normal dan kumpulan yang diberikan ekstrak untuk kedua-dua

    kajian. Kadar perubahan berat organ ratio kepada berat badan untuk ketiga-tiga kajian

    juga menunjukkan tiada signifikan perubahan di antara kumpulan kawalan normal dan

    kumpulan yang diberikan ekstrak. Histopatologi analisis, kajian akut toksisiti

    menunjukkan signifikan perubahan skor nekrosis sel hepar (sedikit ke sederhana) dan

    degenerasi (sangat sedikit) dalam kumpulan dos 2000mg/kg. manakala kajian subakut

    menunjukkan signifikasi perubahan skor (p

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    ACKNOWLEDGEMENTS

    All praises to the Almighty Allah, the most Gracious and Merciful, Who is omnipotent

    and all giving, for affording me the strength and determination to complete this study.

    I would like to express my gratitude to my supervisor Assoc. Prof. Dr. Hazilawati

    Hamzah for her guidance, continued support and encouragement throughout this

    work. I am particularly grateful for my co-supervisors Prof. Dr. Noordin Mohamed

    Mustapha, Assoc. Prof Dr. Norhaizan Md. Esa, Dr. Shanmugavelu Sithambaram and

    Dr. Mohd. Rosly Shaari, for providing me the needed support, good comments and

    invaluable suggestions.

    I wish to express my thanks to the Ministry of Higher Education (MOHE), Malaysia

    Agricultural Research and Development Institute (MARDI) and University Putra

    Malaysia for the scholarship, funding and allowance for supporting this work. I wish

    to mention and thank: Animal house unit staff (AMTREC) and all the staff in the

    Department of Veterinary Pathology & Microbiology, Faculty of Veterinary

    Medicine.

    Acknowledgments are gratefully made to the following people: Farhan, Nadia, Farah,

    Nabila, Amira and other research mates for their contribution in varying degrees of

    this work.

    I wish to express my thanks for my parents and family who always pray, support and

    encourage me both mentally and physically. Finally, I wish to acknowledge with

    thanks all those who, cooperated with me during this work, in all laboratory work and

    who read, reviewed and offered numerous helpful suggestions and corrections